In article
<Pine.A41.3.96.990204142406.62208A-100000 at node5.unix.Virginia.EDU>, Eric
Park <yhp4n at node5.unix.Virginia.EDU> wrote:
>I'm encountering some problems when I try to isolate neutrophils from my
>blood. I'm using a Ficoll-Hypaque solution when I centrifuge the blood.
>This technique works great for several other people but not for me. A lot
>of RBCs are remaining in the neutrophil layer and not settling down to the
>bottom of the tube. Does anyone know why this is happening to my blood?
>>Personal info:
>Blood type is B+ Rh+
>Asian male 22 years old
>>Insights would be helpful since this is a mildly annoying problem in my
>research by begging for volunteers instead of tapping my own blood.
Eric-Are you warming up the Ficoll-Hypaque solution before you layer on
the blood? The solution should be at room temp, and your blood should be
cooled down, before you layer on the blood.
In our lab we separate the neutrophils from the RBC's with Dextran
sedimentation after getting rid of the other WBC's, then use hypotonic
lysis to get rid of any stubborn RBC's.
You may want to try this, but you may want to beg a little blood from
others, too. You can't write a thesis with an N of 1.
Good luck!
Cindy Fuller, PhD, RD
--
C.J. Fuller
<mailto:cjfuller at erickson.uncg.edu>
<mailto:cjfuller at mindspring.com>
